大肠癌中p53基因突变的研究

应用聚合酶链反应(PCR)──单链构型多态性(SSCP)结合银染法对14例大肠癌p53基因的第4、第5─6和第7外显子进行了点突变的研究,结果共检测出6例点突变,而且发现各外显子的突变频率存在差异。另外,利用购自ATCC的两个探针 (p53cDNA探针和pYNZ22探针)对大肠癌中p53基因的杂合性失去进行了研究,在14例大肠癌中共检出6例杂合性丢失。将点突变检测结果同杂合性丢失结果进行比较分析, 并着重探讨了大肠癌中p53基因失活导致肿瘤的作用方式。 Abstract:The exons 4-7 of p53 gene were examined in 14 colorectal Cancer patients by using PCR-SSCP-silver staining method.The results showed 6 cases of point mutation and the mutation frequencies of exons were different from each other.p53 cDNA and pYNZ22 VNTR were used as probes to examine LOH(Loss of heterozygosity)of 14 colorectal cancers.6 cases with LOH were found.The results of present research suggest that mutation and LOH of p53 gene are critical events in the progress and development of Cancer.There were different kinds of inactivation model of p53 gene in the process of development of cancer and transformation of cells.     

罗非鱼TRAF3基因的表达及功能研究

肿瘤坏死因子受体(TNFR)相关因子3(TRAF3)是多种免疫途径中的关键调控因子。从尼罗罗非鱼(Oreochromis niloticus)中克隆获得了TRAF3基因, 命名为OnTRAF3(GeneBank No. MN258118), 该基因包含1个环指结构域、1个锌指结构域、1个卷曲螺旋和MATH结构域。多序列比对表明, OnTRAF3与其他已知的TRAF3蛋白具有高度的相似性, 尤其是MATH结构域。实时荧光定量PCR(qRT-PCR)分析显示, OnTRAF3在各组织中广泛分布, 且在脑、皮肤、肠和鳃中表达量较高。在无乳链球菌诱导后, 多个组织中的OnTRAF3表达量出现了不同程度的上调, 说明OnTRAF3参与了罗非鱼的抗菌免疫应答。亚细胞定位实验显示, OnTRAF3分布在HEK293的细胞质和细胞核中。此外, 荧光素酶报告基因实验结果显示, 野生型(WT)OnTRAF3可显著激活NF-κB信号, 而coiled-coil和MATH 结构域缺失后, 依然能够显著激活NF-κB 活性, 而RING 和Zinc缺失后, 该激活作用则明显减弱, 表明RING 和Zinc 结构域是OnTRAF3在免疫信号通路中行使功能的关键结构域。研究为探索TRAF3在罗非鱼免疫应答中的功能提供了重要的基础。     

Distribution and Significance of Nitric Oxide Synthase in Brain Tissues of Rats Exposed to Deltamethrin Insecticide

The distribution of nitric oxide synthase(NOS)in brain tissues of rats exposed to deltamethrininsecticide has been examined by histochemical NADPH-diaphorase staining techniques on frozen sec-tions.After injection of deltamethrin(12.5mg/kg,i.p.),a reproducible sequence of toxic signs ofhyperexcitability were elicited.The observation and image analysis showed that,within brain sec-tions of rats exposed to deltamethrin,the numbers and the total staining areas of the NOS positiveneurons were greatly increased,especially in cerebral cortex,hippocampal formation and paraventric-ular nucleus.In addition,the density of single neuron and the processes were also increased.The re-sults suggested that deltamethrin may induce the NOS expression or activate the NOS activity.TheNOS activation may involve in the chains responsible for the excitatory neurotoxicities induced bydeltamethrin.     

Re-detectable positive SARS-CoV-2 RNA tests in patients who recovered from COVID-19 with intestinal infection

Dear Editor,Severe acute respiratory syndrome coronavirus 2(SARSCoV-2),a novel coronavirus that causes Coronavirus Disease 2019(COVID-19)(Yang and Wang,2020),has spread to more than 200 countries and infected more than 9,000,000 people as of Jun 2020.Tens of thousands of patients with COVID-19 have recovered and been discharged from hospital.However,there are reports of recovered patients who subsequently tested positive for SARSCoV-2 after discharge(re-detectable positive,RP)(An et al.,2020;Lan et al.,2020),and this has led to increasing focus on the mechanism(s)underlying RP.     

An Integrative Taxonomy of Amphibians of Nepal: An Updated Status and Distribution

Amphibians are declining worldwide.At the same time,each year many species of amphibians are described,indicating that biological diversity on Earth remains poorly understood.This is especially true for mountainous regions,such as the Himalayas,which are also regarded as a biodiversity hotspot.However,it is most likely that the current diversity of amphibians in Nepal is highly underestimated.Amphibian inventories by the past researchers primarily used phenotypic characters as the basis for species identification.However,the application of molecular systematics methods to taxonomic studies has successfully uncovered the taxonomic status of several cryptic taxa.This study used both molecular(16 S rDNA sequences)and morphological comparisons and estimated the diversity of amphibians from Nepal.This study reports the occurrence of 38 species of amphibians based on both molecular and morphological comparisons.Among them,three species were recorded for the first time in Nepal.This study also invalids the taxonomic status of Sphaerotheca paschima.This study highlights the importa nce of integrative taxonomic approaches that help to resolve the current taxonomic ambiguities of Nepalese amphibians.     

The History and Advances of Reversible Terminators Used in New Generations of Sequencing Technology

DNA sequencing using reversible terminators, as one sequencing by synthesis strategy, has garnered a great deal of interest due to its popular application in the second-generation high-throughput DNA sequencing technology. In this review, we provided its history of development, classification, and working mechanism of this technology. We also outlined the screening strategies for DNA polymerases to accommodate the reversible terminators as substrates during polymerization; particularly, we introduced the "REAP" method developed by us. At the end of this review, we discussed current limitations of this approach and provided potential solutions to extend its application.     

Comparative systems biology between human and animal models based on next-generation sequencing methods

Animal models provide myriad benefits to both experimental and clinical research. Unfortunately, in many situations, they fall short of expected results or provide contradictory results. In part, this can be the result of traditional molecular biological approaches that are relatively inefficient in elucidating underlying molecular mechanism. To improve the efficacy of animal models, a technological breakthrough is required. The growing availability and application of the high-throughput methods make systematic comparisons between human and animal models easier to perform. In the present study, we introduce the concept of the comparative systems biology, which we define as "comparisons of biological systems in different states or species used to achieve an integrated understanding of life forms with all their characteristic complexity of interactions at multiple levels". Furthermore, we discuss the applications of RNA-seq and ChIP-seq technologies to comparative systems biology between human and animal models and assess the potential applications for this approach in the future studies.     

Secretory/releasing proteome-based identification of plasma biomarkers in HBV-associated hepatocellular carcinoma

For successful therapy, hepatocellular carcinoma (HCC) must be detected at an early stage. Herein, we used a proteomic approach to analyze the secretory/releasing proteome of HCC tissues to identify plasma biomarkers. Serum-free conditioned media (CM) were collected from primary cultures of cancerous tissues and surrounding noncancerous tissues. Proteomic analysis of the CM proteins permitted the identification of 1365 proteins. The enriched molecular functions and biological processes of the CM proteins, such as hydrolase activity and catabolic processes, were consistent with the liver being the most important metabolic organ. Moreover, 19% of the proteins were characterized as extracellular or membrane-bound. For validation, secretory proteins involved in transforming growth factor-β signaling pathways were validated in plasma samples. Alphafetoprotein (AFP), metalloproteinase (MMP)1, osteopontin (OPN), and pregnancy-specific beta-1-glycoprotein (PSG)9 were significantly increased in HCC patients. The overall performance of MMP1 and OPN in the diagnosis of HCC remained greater than that of AFP. In addition, this study represents the first report of MMP1 as a biomarker with a higher sensitivity and specificity than AFP. Thus, this study provides a valuable resource of the HCC secretome with the potential to investigate serological biomarkers. MMP1 and OPN could be used as novel biomarkers for the early detection of HCC and to improve the sensitivity of biomarkers compared with AFP.